The goal of the work proposed in this application is to obtain a set of cosmid clones which overlap and span in a continuous map the major portion of human chromosome 11. For the past two years, our research group has been involved in obtaining saturating collections of cosmid clones for 11q13- 11qter, a region including about 1/3 of chromosome 11, and in developing new methods for determining overlaps between the cosmids. The work proposed in this application will apply several different strategies and technologies to the analysis of the entire chromosome, concentrating on the additional 2/3 not represented in our current collection. This will include: (1) production of cosmid libraries from DNA isolated from flow purified chromosome 11 (2) archiving these clones in 96-well microtitre plates and organizing cosmids in two- or three-dimensional arrays (3) determining overlaps between cosmids in the collection for the construction of contigs using (i) multidimensional cosmid multiplex analysis and (ii) detailed restriction mapping of a large portion of the set by automated DNA preparation and digestion, (4) locating mapping landmarks using synthetic oligonucleotide probes, (5) detecting linking clones containing rare restriction sites by automated restriction digestion analysis, (6) mapping landmark clones to chromosomal locations by high resolution in situ hybridization, (7) linking clustered contigs and linking clones by pulsed field gel electrophoresis, and (8) filling in the gaps in the map using a combination of directed cosmid "walking", pool analysis using end clones, and yeast artificial chromosomes. The studies proposed here will result in a detailed physical map and set of reference clones for human chromosome 11. It will thus serve as the basis for the localization of important disease genes, such as those responsible for ataxia telangiectasia, multiple endocrine neoplasia Type 1, tuberous sclerosis, peripheral neuroepithelioma, and others. In addition, it will allow the detection and analysis of genes possibly important to human biology and development such as genes containing known structural motifs, genes syntenic to murine developmental mutations (i.e., NOD) and genes involved in chromosomal translocations important in malignancies (i.e., Acute leukemias and Ewing's sarcoma). This collection will also provide template sets for possible eventual DNA sequencing of selected portions of chromosome 11.